Canine TNF-alpha Antibody

(1 citations)   
  • Species Reactivity
    Canine
  • Specificity
    Detects canine TNF-alpha in direct ELISAs. In direct ELISAs, 25-100% cross-reactivity with recombinant human TNF-alpha, recombinant porcine TNF-alpha, and recombinant rhesus macaque TNF-alpha is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 236812
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant canine TNF-alpha
    Val77-Leu233
    Accession # P51742
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Immunocytochemistry
    8-25 µg/mL
    See below
    • Canine TNF-alpha Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Canine TNF‑ alpha Antibody (Catalog # MAB1507)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Canine TNF‑ alpha Biotinylated Antibody (Catalog # BAF1507)
  • ELISA Standard
     
    Recombinant Canine TNF-alpha Protein (Catalog # 1507-CT)
  • Neutralization
    Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. The Neutralization Dose (ND50) is typically 1.25-5.0 µg/mL in the presence of 10 ng/mL Recombinant Canine TNF‑ alpha and 1 µg/mL actinomycin D.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody. Recombinant Canine TNF‑ alpha (Catalog #
1507-CT) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line). Cytotoxicity elicited by Recombinant Canine TNF‑ alpha (10 ng/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Canine TNF‑ alpha Monoclonal Antibody (Catalog # MAB1507). The ND50 is typically
1.25-5.0 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL).
Immunocytochemistry
TNF‑ alpha in Canine PBMCs. TNF‑ alpha was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) untreated or treated with calcium ionomycin and PMA using Mouse Anti-Canine TNF‑ alpha Monoclonal Antibody (Catalog # MAB1507) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell secretion. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-alpha is produced by a wide variety of immune and epithelial cell types (1, 2). Canine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). Within the ECD, canine TNF-alpha shares
84‑94% aa sequence identity with equine, feline, human, porcine, and rhesus and 69-77% with bovine, cotton rat, mouse, and rat with TNF-alpha. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer (4). Ligation of this complex induces reverse signaling that promotes lymphocyte co-stimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in the development of several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11).

  • References:
    1. Idriss, H.T. and J.H. Naismith (2000) Microsc. Res. Tech. 50:184.
    2. Hehlgans, T. and K. Pfeffer (2005) Immunology 115:1.
    3. Zucker, K. et al. (1994) Lymphokine Cytokine Res. 13:191.
    4. Tang, P. et al. (1996) Biochemistry 35:8216.
    5. Eissner G. et al. (2004) Cytokine Growth Factor Rev. 15:353.
    6. Black, R.A. et al. (1997) Nature 385:729.
    7. Moss, M.L. et al. (1997) Nature 385:733.
    8. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
    9. Clark, I.A. (2007) Cytokine Growth Factor Rev. 18:335.
    10. Romanatto, T. et al. (2007) Peptides 28:1050.
    11. Hector, J. et al. (2007) Horm. Metab. Res. 39:250.
  • Long Name:
    Tumor Necrosis Factor alpha
  • Entrez Gene IDs:
    7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)
  • Alternate Names:
    APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; tnfa; tnf-a; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Expression of Toll-like receptors 2 and 9 in cells of dog jejunum and colon naturally infected with Leishmania infantum.
    Authors: Figueiredo M, Amorim I, Pinto A, Barbosa V, Pinheiro L, Deoti B, Faria A, Tafuri W
    BMC Immunol, 2013;14(0):22.
    Species: Canine
    Sample Type: Tissue Homogenates
    Application: ELISA Development
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Mouse IgG Antibody

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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

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Mouse IgG VisUCyte HRP Polymer Antibody

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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

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